CAJ | 학술논문

核胞浆转运蛋白类%氧化性应激%基因表达%反应元件%博来霉素/投药和剂量%抗药性%肿瘤%肺肿瘤%衔接蛋白质类%膜泡运输%有机铂化合物覈胞漿轉運蛋白類%氧化性應激%基因錶達%反應元件%博來黴素/投藥和劑量%抗藥性%腫瘤%肺腫瘤%銜接蛋白質類%膜泡運輸%有機鉑化閤物
핵포장전운단백류%양화성응격%기인표체%반응원건%박래매소/투약화제량%항약성%종류%폐종류%함접단백질류%막포운수%유궤박화합물
Karyopherins%Oxidative stress%Gene expression%Response elements%Bleomycin/admin%Drug resisitance,neoplasm%Lung neoplasms%Adaptor proteins,vesicular transport%Organoplatinum compounds

目的:建立野生型Keap1过表达的H460细胞株降表达Nrf2,检测Nrf2-ARE信号通路对肿瘤细胞耐药性的影响.方法:H460细胞转染mKeap1-pEGFP后经过长期筛选得到稳定表达Keap1蛋白质的细胞株,通过Western blotting和荧光定量PCR的方法进行检测;并通过多次继代培养,细胞株H460-N5稳定表达mKeap1,Nrf2及其调控基因均显著降表达;用MTS法检测抗癌药物奥沙利铂、阿霉素和依托泊苷对细胞增殖抑制率.结果:H460细胞转染mKeap1-pEGFP后筛选建立Nrf2降表达的稳定细胞株H460-N5.MTS数据显示,与对照组相比,抗癌药物奥沙利铂、阿霉素和依托泊苷对H460-N5细胞的抗增殖作用更显著.当奥沙利铂和依托泊苷分别在93 μmol/L和100 μmol/L浓度作用于对照组细胞株H460-N0时,药物作用接近半数抑制率(IC_(50));而在H460-N5细胞株中,两种抗癌药物的IC_(50)分别为42 μmol/L和30 μmol/L.阿霉素对H460-N0细胞的IC_(50)>3 mg/L,而对H460-N5细胞的IC_(50)约为2 mg/L.结论:Keap1过表达的H460-N5细胞Nrf2及调控基因显著降低并增敏抗癌药物奥沙利铂、阿霉素和依托泊苷对肿瘤细胞的增殖抑制作用.
목적:건립야생형Keap1과표체적H460세포주강표체Nrf2,검측Nrf2-ARE신호통로대종류세포내약성적영향.방법:H460세포전염mKeap1-pEGFP후경과장기사선득도은정표체Keap1단백질적세포주,통과Western blotting화형광정량PCR적방법진행검측;병통과다차계대배양,세포주H460-N5은정표체mKeap1,Nrf2급기조공기인균현저강표체;용MTS법검측항암약물오사리박、아매소화의탁박감대세포증식억제솔.결과:H460세포전염mKeap1-pEGFP후사선건립Nrf2강표체적은정세포주H460-N5.MTS수거현시,여대조조상비,항암약물오사리박、아매소화의탁박감대H460-N5세포적항증식작용경현저.당오사리박화의탁박감분별재93 μmol/L화100 μmol/L농도작용우대조조세포주H460-N0시,약물작용접근반수억제솔(IC_(50));이재H460-N5세포주중,량충항암약물적IC_(50)분별위42 μmol/L화30 μmol/L.아매소대H460-N0세포적IC_(50)>3 mg/L,이대H460-N5세포적IC_(50)약위2 mg/L.결론:Keap1과표체적H460-N5세포Nrf2급조공기인현저강저병증민항암약물오사리박、아매소화의탁박감대종류세포적증식억제작용.
Objective: To maked a Nrf2 down-regulated cell line by over-expressing Keap1 in H460 cells to study the role of Nrf2 in drug resistance. Methods: Transfecting H460 cells with mKeap1-pEGFP and screennig for Keap1 expressing clones by Western blotting with antibodies against Nrf2,HO-1,NQO1 and AKR1C.The cell line with Keap1 over-expression was further confirmed by real-time PCR.The cytotoxicity of H460-N5 to anti-cancer drugs was evaluated by MTS assay. Results: MTS assay results showed the enhanced cytotoxicity of anticancer drugs (Oxaliplatin,Doxorubicin and Etopside) to the H460 cell line with keap1 overexpression compared to the control cell line.In H460-N0 cells,the IC_(50) values of Oxaliplation and Etopside were 93 μmol/L and 100 μmol/L respectively whereas the IC_(50) values of the two drugs were 42 μmol/L and 30 μmol/L correspondingly in H460-N5 cells.The IC_(50) value of Doxorubicin to H460-N0 cell was above 3 mg/L,but that to H460-N5 cell was about 2 mg/L. Conclusions: A Nrf2 down-regulated cell line H460-N5 and a control cell line with GFP over-expression have been identified.Down-regulation of Nrf2 enhanced the cytotoxicity of Oxaliplatin,Doxorubicin and Etopside.